Terapia Celular y Reparación del Sistema Nervioso Central
Profesor Gustavo A Moviglia Dr. en Medicina. Director del CIITT
Universidad Maimónides – Buenos Aires - Argentina
Terapia con Células
El uso de células vivas, proce-sadas ex – vivo y reintroducidas en un organismo humano con el objeto de curar, mitigar, prevenir o diagnosticar una enfermedad o condición patológica.
E. Donnall Thomas Desarrollo desde el
año 1955 en animales
y humanos (1957) el
transplante de
Médula Ósea. Por
este motivo se le
otorgó el Premio
Nobel de Fisiología y
Medicina en el Año
1990
Transplantes de células hematopoyéticas
Laflamme et al. Circ Res. 2002;90:634-640.
Lecciones del Trasplante de Células Hematopoyéticas
1.- Las células, implantadas por transfusión venosa, superan el filtro pulmonar y se dirigen a la despoblada médula ósea. Se implantan donde el organismo las necesita y les permite.
2.- Su actuar no es instantáneo sino que se despliega durante los día subsiguientes al implante.
3.- Las células y el organismo receptor deben aceptarse mutuamente. De su correcta recepción, por parte del paciente, depende el que estas células puedan comenzar a producir plaquetas, glóbulos rojos y glóbulos blancos.
Graft Versus Host Diseases
Efecto Injerto Contra Leucemia
Evolución de la Lesión del IAM
Necrosis - Apoptosis
Inflamación
Angiogenesis - Vascularización
Regeneración
0 4 2 6 8 10 14 12 16 18 20
Días
Cicatrización
Ventana Terapéutica
1 día
40° día 15° día 7°-10° día
2°-3° día
Evolución del IAM
Puntos de Vista Parciales
Petrovic-Djergovic et al. Circ Res. 2016;119:142-158
Th1 Inflammation Acts Synergistic with Cellular Therapy
Popa-Wagner et al. Neural
Plasticity Volume 2015,
Article ID 839638, 7 pages
Th2 Inflammation as Detrimental for Neurogenesis
Factores que Promueven La Reparación Tisular
Células Pro-reparadoras (Inductoras y Estructurales)
Apoyadas sobre una matriz extracelular apropiada
Embebidas en un medio humoral adecua-do
Exigido funcionalmente en la justa medida
Protective Autoimmunity Induced by Lymphocytes Procures SCI Recovery.
MBP ETC
GFAP Stain
Injured Site Attracts Activated T Cells
TCR Stain
D.L. Hirschberg et al. Journal of Neuroimmunology. 1998; 89:88–96
Activated T Cell Mediated Repair
Hauben E et al. J Neurosci 2003; 25: 8808-19
Protective Autoimmunity induced by self antigen (peptide) vaccination
Barouch & Schwartz, FASEB J. 2002; 16:1304
Barouch & Schwartz, FASEB J. 2002; 16:1304
Secretion of Neurotrophins by T Cells after MBP Vaccination
BDNF NT-3
Therapeutic Window for Activated T Cell Mediated Repair
Yoles et al. J Neurosci. 2001, 21(11):3740-3743
Patient
MSC Effecter Cells
Co-Culture
MSC differentiation into NPC
MSC After 48 hr of Co Incubation with EC against
Polipeptides from Hydrolyzed Cow Brain
Control MSC Culture MSC + ETC 48 hr Culture
Lymphocytes against hydrolyzed bovine brain induce the differentiation of BM-MSC to NPC
Conditioned Medium
from Activated Effector
Cells
Activated Effector
Cells Only
Non Activated Effector
Cells and Hydrolyzed
Bovine Brain
+
+
+
MSC
MSC
MSC
NPC
MSC
MSC
MSC After 48 hr of Co Incubation with activated
EC against Polypeptides of Cerebrolysin
Lymphocytes against Cerebrolysin induce the differentiation of Ad-MSC to NPC
Electric Activity of Differentiated MSC
MC
RGC
RPE
Rods
A
MSC
B
D C
RPC
0 Hr after Co culture with ETC 24 Hr after Co culture with ETC
72 Hr after Co culture with ETC 72 Hr after Co culture with ETC
EC against hydrolyzed swine retina induce differentiation of Ad-MSC to RPC
a b c
Confocal microscopy analysis of the cultured cells. a GFAP-positive cells. b Bestrophin 2-positive cells.
c Merge a and b image. d Tubulin beta III-positive cells. e OPN1 SW-positive cells
a
e d
c b
Lymphocytes against hydrolyzed swine pancreatic islets
induce differentiation of Ad-MSC to insulin and glucagon
producer cells
MSC 48 Hr MSC+ETC+PIM Culture Immunoperoxidase Insulin Staining
MSC 48 Hr MSC+ETC+PIM Culture Immunoperoxidase Glucagon
Staining
Insulin and Glucagon Secreting Islet
elaborated from Patient adMSC
Taguchi A et al. Stem Cells Dev. 2015 ;
24(19):2207-18
BMMNC at 7 days post Stroke - Clinical
Trial Phase 1-2
Taguchi A et al. Stem Cells Dev. 2015 ;
24(19):2207-18
BMMNC at 7 days post Stroke - Clinical
Trial Phase 1-2
Zhang et al. BMC Neurosci (2015) 16:56
Cellular Therapy and Rehabilitation
Clinical Differences between Acute and Chronic Spinal
Cord Injured Patients Associated to Therapeutic Strategy
Chronic Lesion Therapy Element Acute Lesion
• Large Blood Supply • Poor Blood Supply • Mesenchymal Stroma
Cell (adipose) IA Infusion
• Acute (Th1) Inflammatory
Reaction
• Chronic (Th2)
Inflammatory Reaction • Effector Cell IV Infusion
• Presence of Inside and
Outside Stem Cells.
• Differentiated Stem Cell
IA Infusion
• Yes Reversible Neural
Engram Disruption.
• No Reversible Neural
Engram Disruption. • Neural rehabilitation
• Absence of Inside and
Outside Stem Cells.
T12 ASIA A/Frankel A, Flaccid paraplegic, 10 Years Evolution,
RESULTS AFTER 30 MONTHS OF TREATMENT
Tibialis Anterior
Muscle L4-S2
Spontaneous Recovery of Muscle innervation of a Frankel A SCI Patient only will be
achieved until 2.5 years after the accident and results no will go further than 3
segments below of the last preserved spinal cord segment.
Rational of the MED Therapeutic Approach for
Optic Nerve and Retina Damage
Restoring the disrupted (choroidal) blood vessels
through Intra Ophthalmic Artery Infusion of cultured
autologous a-MSC
Modulating the Inflammatory Reaction through Intra
Venous Auto Aggressive Effector T Cell Infusion.
(ETC)
Restoring the Supply of Specific Stem Cells through
Intra Artery Infusion of Differentiated MSC into Retina
Progenitor Cell. (DSC)
Intra Artery Infusion
Treatment Results of Optic
Nerve Lesion
Pretreatment Optic Nerve
Tractography
Optic Evoqued Potentials of the Right Optic Nerve
Wave Pre Treatment Post Treatment
N1 171 60
P1 200 119
N2 224 167
Electroretinogram of the Right Retina
Wave Pre Treatment Post Treatment
a 12,9 11,7
b 49,2 49,2
Optic Coercive Tomography
(OCT)
Comparación Imágenes de OCT e
Histología de la Retina
Pathogenic process of Age Related Macular Degeneration (AMD)
Age Related Macula Degeneration (AMD)
Pre Treatment 3 months after Treatment
Pre Treatment 3 months after Treatment
Preliminary Results
After three months of the cellular treatment of a Patient with only Optic Nerve Damage: Recovery of Optic Evoked Potential (OEP).
Persistence of the normal values of the Electroretinogram (ERG)
After three months of the Cellular treatment of two Chronic Blind AMD patients, in the OCT and Digital Biopsy Analysis, was observed: 4 times Enlargement of the Chronic Atrophied Choroidal
Vessels.
Decreasing of the sub retina and intra retina edema areas.
Increasing up to 20 time of the number of Retinal Pigmented Epithelium cells (RPE) in the previous depopulated areas.
Retina width Increasing of 20 to 25% in previous atrophied areas.
Conclusions
Optic Evoked Potentials (OEP) and Electroretinogram (ORG) Registers as Well as OCT and Digital Biopsy Analysis may be effective, accurate and quantitative methodol-ogies to objectively evaluate Safety and Efficacy of Cellular therapies applied for Optic Nerve, Retina and CNS pathologies.
Clinical results obtained in the Ophthalmologic setting seem be similar to the ones obtained in the treatment of Spinal Cord Injury. This fact may shear light on the Mode of action of MED therapy program.
Clinical trials are in progress to asset the above preliminary conclusions.